rabbit anti-pgk  (Thermo Fisher)

Journal: bioRxiv

Article Title: Mitochondrial targeting of glycolysis in a major lineage of eukaryotes

doi: 10.1101/257790

Figure Lengend Snippet: Phaeodactylum tricornutum contains, similar to some other stramenopiles, multiple isoforms for the C3 part of glycolysis. The localization for all isoforms was tested via GFP-fusion constructs. A “pre” suffix means that the predicted targeting signal was used; if the suffix is missing the full length of the respective sequence was fused to GFP. The number is the JGI Protein ID and the result of each localization is mentioned. For the corresponding amino acid sequences used for GFP targeting see Supplementary File 2. A star (*) marks images were a maximum intensity projection from a Z-Stack was used. Unclear indicates localization not possible to identify. TPI = Triosephosphate isomerase, GAPDH = Glyceraldehyde-3-phosphate dehydrogenase, PGK = Phosphoglycerate kinase, PGM = Phosphoglycerate mutase, ENO = Enolase, PK = Pyruvate kinase. DIC, Differential interference contrast microscopy. Chl, Chlorophyll a autofluorescence. GFP, Green fluorescent protein. Chl+GFP, Merged imaged showing the discrete localization of GFP compared to Chlorophyll autoflourescence. For the corresponding amino acid sequences used for GFP targeting, see Supplementary File 2. Scale bar 5 μm.

Article Snippet: Primary antibodies: Rabbit, anti-PGK; Guinea Pig, anti-TPI-GAPDH (Eurogentec, Seraing, Belgium) were used at a dilution of 1:500 and 1:300 in 2% BSA in PBS, 0.1% triton X-100, respectively.

Techniques: Construct, Sequencing, Microscopy